Shouting at the ocean with pebbles in my mouth since 2008. The subjects of this blog include forensics, the war-on-terror detainees, the Duke lacrosse case, the Knox/Sollecito case, and the academic world as it intersects the political. It will sometimes examine issues of particular interest to Wilmington, NC and the University of North Carolina at Wilmington.

Sunday, May 9, 2010

Secondary transfer of DNA and DNA contamination

Part XV in the Knox/Sollecito case

IntroductionThis post returns to the subject of forensic DNA contamination. It will define primary and secondary transfer of DNA and explain how they are related to contamination. It also treats clerical errors and the need for independent review of the forensic DNA data. It reconsiders the lessons of the Leskie case. These concepts will be applied to some of the DNA forensics in the murder of Meredith Kercher, especially as it relates to the evidence against Amanda Knox and Raffaele Sollecito.

DNA contaminationPeople sometimes confuse DNA contamination with secondary transfer of DNA. Let us define these terms and give a few examples. In Forensic DNA Typing, p. 152, John Butler wrote, “Contamination implies the accidental transfer of DNA.” Implicit in his definition is that the transfer happened after the evidence was collected. In An Introduction to Forensic DNA Analysis, p. 14, Norah Rudin and Keith Inman “define contamination as the inadvertent addition of an individual’s physiological material or DNA during or after collection of the sample as evidence…A contaminated sample is one in which the material was deposited during collection, preservation, handling, or analysis.”

Primary transfer of DNAAccording to forensic scientist Suzanna Ryan, “Primary transfer is described as the transfer of DNA from an individual to an item.” Ryan summarized some of the factors that contribute to how much DNA is transferred: “Through studies by Ladd, et al and others, many in the forensic community believe that some individuals just naturally shed more cells when handling an item (‘good’ shedders) than others (‘poor’ shedders). Other factors including the substrate being handled (rougher items collect more DNA), the time since the individual last washed their hands, how nervous the person is (nervousness can lead to increased sweating), and how often they touch their mouth, eyes, hair, face or other body parts (thus gathering DNA on their hands) play a heavy role in whether a DNA profile will be obtained through simply touching an item.” One presumes that rougher items cause more cells to transfer than smoother items, but Ms. Ryan did not give specific examples.

Secondary transfer of DNASuzanna Ryan wrote, “In secondary transfer, there is no direct contact between a person and an object. It is instead transferred through an intermediary. This could be another person or another object. An example—expanding upon the same knife example as above—is as follows: Person A shakes Person B's hand. Person B touches the handle of a knife. Secondary transfer theory would allow that Person A's DNA could be transferred first to Person B's hand and second to the knife handle. This would mean that even though Person A never actually touched the knife handle, his DNA could be present on it.”

Secondary transfer at the scene and the labSuzanna Ryan continued, “At the laboratory, the analysts are careful to wear gloves when handling evidence items. Great care is generally taken to clean scissors, tweezers, and other utensils between testing items. But what about items that are not cleaned as regularly? For example, is it possible to transfer DNA from an item of evidence to a ruler, and then when the next item of evidence is examined and photographed, could the DNA transfer again from the ruler to this item? A 2006 study by Poy and van Oorschot showed an example of secondary transfer when a mixed DNA profile was found on a swab taken from an examination magnifying lamp. This profile was searched in the lab's database and a match was found with a case that had been worked on the bench-top with the magnifying lamp. It was determined that DNA was transferred from the item being examined to the analyst's gloves and then onto the top of the magnifying lamp.”

This incident should be considered both secondary transfer of DNA (the item transferred DNA to the glove, which transferred it to the lamp) and DNA contamination because it happened in the analysis lab. This incident also illustrates how easy accidental DNA transfer can be in a lab. DNA contamination occurs when an analyst transferred his or her own DNA onto an object, but this would be classed as primary transfer.

Sometimes DNA contamination occurs in ways that do not involve the item of evidence. DNA profiles have been observed because an item of labware (plastic tubes, cotton buds, etc.) was contaminated with DNA from a person who helped to manufacture the item. The chemical reagents and equipment needed to perform the polymerase chain reaction, the DNA amplification step, are occasionally contaminated. These are examples of contamination that are due neither to primary nor to secondary transfer.

Primary and secondary transfer when several people handle an objectIn the journal Nature, 387, p. 767 (1997), R. van Oorschot and M. Jones wrote, "Objects handled by many individuals all produced profiles with multiple alleles of varying intensity. To determine the effect of multiple handlers, we exchanged polypropylene tubes between individuals (2 or 3, 10 min each) with different genotypes. Although the material left by the last holder was usually present on the tube, that of previous holders was also retrieved to varying extents. The strongest profile obtained was not always that of the person who last held the object, but was dependent on the individual. We regularly observed profiles of previous holders of a tube from swabs of hands involved in these exchanges, showing that in some cases material from which DNA can be retrieved is transferred from object to hand (secondary transfer).” This article was the first one of which I am aware to document that DNA profiles could be recovered from fingerprints. This may also be the first one to use the term “secondary transfer.”

Primary and secondary transfer of DNA to the bra claspSome have argued that the amount of Raffaele Sollecito’s DNA on the clasp can only have been deposited through primary (direct transfer), not contamination or secondary transfer. This argument is false on several grounds. It ignores that that the amount of DNA in primary transfer depends upon several factors (see above). It ignores that contamination can produce strong profiles that analysts have mistaken for genuine ones (see below). It ignores the fact that Mr. Sollecito’s DNA profile on the clasp was relatively weak in intensity, roughly 200 RFUs. Perhaps most fundamentally, this argument runs counter to a principle of DNA forensics. As part of his analysis of the Leskie case (see below), forensic scientist Dan Krane wrote, “In fact, it is quite uncommon for DNA tests themselves to say anything about the circumstances (or even the time frame) associated with the transfer.” Jason Gilder wrote in an email to me, “One of the standard axioms of DNA typing is, the presence of a DNA profile says nothing about the time frame or the circumstances under which DNA was transferred to that item. Contamination falls into is one such instance of an issue that cannot be identified by the electropherogram alone (unless you are dealing with a control sample or other known profile).”

On the other hand, an anonymous commenter on a discussion board offered this hypothetical scenario for secondary transfer of Mr. Sollecito’s DNA: “For example, let's say that DNA from Meredith, Amanda and Raffaele was on the hand towels Rudy took from the bathroom (highly probable, since they'd all used the bathroom in the days before the murder). Drying your hands on a hand towel is exactly the kind of rubbing action that would shed DNA. If Rudy had trodden on one of the towels, then trodden on the bra clasp (one of the hooks was deformed, of course) that could have transferred DNA from the towels to the hook, with no direct contact from Raffaele necessary. In turn, Raffaele's DNA was more easily identified because they could use the more sensitive tests for the Y chromosome, whereas the other DNA would have been mixed female and thus tricky to correctly identify.” This is a speculative scenario, but it is not the only kind of secondary transfer that one could imagine. Sollecito’s DNA was likely on the door to Meredith’s bedroom, for example, because he tried to break the door down.Clerical errors in DNA forensicsPreviously we have explored why the electronic files are important. However, simply having an independent expert review the case file can catch some types of errors even without the electronic files. DNA forensic consultant Norah Rudin encountered a case where two reference samples were switched, due to a clerical error. Lazaro Sotolusson was held for a year and would have been tried for rape involving juveniles, facing multiple life sentences. In a preliminary hearing, one of the female victims identified Sotolusson as her attacker, possibly a case of a mistaken eyewitness. Dr. Rudin examined handwritten notes and computer files (not the sort of files one uses to create a DNA electropherogram, such as a .fsa file) and concluded that the lab had switched the two samples. The lab retested samples from Mr. Sotolusson and Joseph Coppola and confirmed the error. Rudin stated to a reporter, “If I hadn't looked at the original data, I likely wouldn't have found the mistake. That's one of the reasons that I always look at the original data… I believe every case needs to be reviewed by an independent analyst.” (emphasis added)

Errors of a similar kind have occurred elsewhere. Professor William Thompson wrote, “I recently reviewed the corrective action file for an accredited California laboratory operated by the District Attorney’s Office of Kern County (Bakersfield). Although this is a relatively small laboratory that processes a low volume of samples (probably fewer than 1,000 per year), during an 18-month period, it documented multiple instances in which (blank) control samples were positive for DNA, an instance in which a mother’s reference sample was contaminated with DNA from her child, several instances in which samples were accidentally switched or mislabeled, an instance in which an analyst’s DNA contaminated samples, an instance in which DNA extracted from two different samples was accidentally combined into the same tube, falsely creating a mixed sample, and an instance in which a suspect tested twice did not match himself (probably due to another sample-labeling error).” (emphasis added)

Whether or not an independent analysis would have caught possible clerical errors here is an open question. The Knox defense team asked for an independent analysis of all of the forensic data, but the judge denied the request. As long as human beings are part of the testing process, clerical errors are likely to happen at some frequency. This factor and the possibility of contamination ought to be considered when one ponders the astronomical odds that are sometimes used when describing a particular DNA result.The Leskie case and its relationship to the knife and bra clasp as evidenceJaidyn Leskie was a toddler who was murdered in Australia over a decade ago. A DNA profile belonging to a Ms. P. was found on two pieces of Jaidyn’s clothing, a bib and some track pants. Ms. P’s profile had varying intensities at different loci, suggesting sample degradation. However, her profile was quite strong in some loci, having peak intensities of over 1000 RFUs. Another example of contamination occurred in State of Illinois v. Sean Evans (2003). In this example a positive control was contaminated with a sample containing a DNA ladder, a series of DNA fragments of increasing length that might be used as a calibration standard. Note that the intensity of the fragments making up the ladder is 500-1500 RFU, similar to a typical evidence sample. The unusual nature of the ladder DNA helps to show that contamination occurred, but this is seldom true for other kinds of contamination.

Ms P’s DNA profile was ultimately attributed to contamination partially on the basis that Ms. P lived hundreds of miles away from the crime and never left her village. Yet she could have lived next door to the Leskie family, and contamination would still be a likely explanation for finding her DNA on the toddler’s clothing. Jaidyn’s body was found submerged in water for months. These conditions are antithetical for the preservation of DNA evidence. Significantly, none of Jaidyn’s DNA was recovered from his clothing. This should have been a reason to suspect contamination on its own.

The bloodless kitchen knife in the Knox/Sollecito case is similar to Jaidyn’s clothing in that no human DNA was likely to have been on the blade at the time of collection at any section that tested negative for blood; therefore, the DNA was most likely deposited in the lab. The Leskie case was one in which the electronic data files were particularly helpful. The Knox/Sollecito case would also benefit by independent examination of these files. For example, one could use the .fsa files to examine other samples along the knife blade and handle using the same threshold value for all samples. One might find profiles belonging to other individuals, and this would strengthen the argument that DNA on the knife arose from contamination or secondary transfer.

Could contamination have occurred in the Leskie case? The evidence with Ms P’s DNA was examined within a few days the evidence from the Leskie case, and Dr. Krane noted “even if evidence samples from the two cases did not come into direct physical contact, secondary transfer (by way of common contact with a surface or implement) of biological material from the condom or articles of Ms. P’s clothing in the rape investigation to the bib and track pants could have occurred.” The lab in question had argued that contamination was unlikely on at least two grounds, one of which was the lack of DNA from Ms P showing up in other profiles. Dr. Krane agreed that “no other samples including negative controls associated with both investigations display indications of Ms. P’s allele’s being present.” However, he rebutted the lab’s point by stating, “there is no good reason to expect that contamination would uniformly affect all samples if and when it does occur. In much the same way, Salmonella-tainted meat placed on one part of a countertop at the beginning of food preparation does not mean that all food subsequently prepared for the same (or even a later) meal will be similarly tainted – just those items that also come in contact with the same portion of the countertop can be reasonably expected to have a chance of being tainted.” Although the amplification of DNA through use of the polymerase chain reaction (PCR) creates the possibility of contamination via the amplified DNA, Dr. Krane did not think it is as likely in this case as secondary transfer.

There is one more point with respect to the bra clasp that needs to be addressed. It is sometimes argued that Mr. Sollecito’s DNA could not have arrived on the clasp by contamination because his DNA was found on no other items except for a cigarette butt. This is essentially the reasoning of Judge Micheli, who oversaw Rudy Guede’s trial. From the Wikipedia article on the murder of Meredith Kercher, “The judge at the trial of Guede acknowledged that the DNA sample was considered small, but described the claim of contamination at the laboratory as making ‘no sense’, since there was no material from which such contamination could have come, and so ‘the risk would have been the loss of traces found there, not the risk of somehow discovering new traces.’” This argument is false for at least two reasons. It ignores the possibility of contamination from Mr. Sollecito’s DNA in the lab, and it ignores the empirical facts of the Leskie case, where Ms. P’s DNA did not show up in other items or controls.

SummaryOne way to distinguish between secondary transfer and contamination is to say that the difference between primary and secondary transfer depends upon how the DNA was deposited on the evidence item, but contamination depends upon when the DNA was deposited: if the DNA were deposited before the item was collected as evidence, it would not be contamination. We have also seen that contaminating DNA does not evince a difference in intensity that would allow it to be distinguished from DNA arising from primary or secondary transfer before the item was collected as evidence. Clerical mix-ups are another source of error in DNA forensics, and catching them is one of many reasons to welcome independent reviews of the results. Raffaele Sollecito’s DNA could have arisen on the bra clasp from contamination or secondary transfer, as well as primary transfer. Meredith Kercher’s weak, dubious DNA profile on the knife is almost certain to have arisen from contamination.

Updates 1/13/2012 and 1/19/02

I corrected Suzanna Ryan's name from Suzanne, and I fixed a link to one of her articles. I added a quotation mark and fixed a link to William C. Thompson's article.

137 comments:

Rose
said...

Wow,It is unfortunate that Meredith's DNA somehow got transferred to that knife and not any of the lab technicians, forensic technicians or other people's DNA they may have been handling in that lab, including DNA from other cases they were working. What are the odds that it would be just happen to be Meredith's.

It is doubly unfortunate that Raffaele's DNA ended up on the bra clasp as well. Just consider what the odds of that would be. Nope, I am convinced it is that rascal Fate again out to get Raffaele and Amanda in hot water. It could be nothing else.

I apologize for the heavy sarcasm in my above post. The point I was trying to make is this:

I really see the DNA results as having 3 probabilities. Either the results on both are genuine, or the results on one of them is genuine and the other the result of cross contamination. The third probability is that both pieces of DNA evidence were deliberately falsified. The possibility that both items are a result of cross contamination that just happen to be contaminated by the two suspects and murder victim are slim in the extreme. If I were a Vegas guy I would put that far, far down on the list. The possibility of deliberate falsification or legitimate results are much higher than both items being cross contaminated this way.

I think that the knife may have been contaminated in the lab. However, I think that both secondary transfer and contamination during collection are possibilities for the clasp. I would say that the people who want to use the bra clasp as evidence against Sollecito are obliged to say how the DNA from three other people got there.

One more point on contamination. In the “contamination” link I provided there is a case from Newfoundland. The Globe and Mail story is called “The Dark Side of DNA,” and the author is Kirk Makin. DNA from a murder victim was found on a wedding ring belonging to Gregory Turner. Mr. Makin wrote, “The technician conceded at Mr. Turner's 2001 trial that she had also contaminated evidence in two previous cases. In another disturbing twist, it emerged that she had mistakenly contaminated Mr. Turner's ring with her own DNA, causing police to waste considerable time on a futile search for a presumed accomplice.” The take-home message for me is that an underperforming technician can beat the odds, shall we say.

As argued by you and others, the prosecution should turn over all data, including the electronic data files, to the defenses. I don't see how a court of law in a civilized nation, whether it be Italy, USA, etc., could justify such a refusal.

Will the appeals court allow and ask for an independent review of the DNA evidence? If the DNA evidence was as unassailable as the prosecution contends, they shouldn't have any problem with an independent review.

It's interesting that at the first trial independent experts offered testimony - about the knife wounds, sexual violence, and the murder dynamic - that didn't support the prosecution's theory.

It really is up to the prosecution to prove the DNA collection and testing is reliable...at least it should be up to the prosecution...and probably is in most places.

The total, entrenched, resistance in Perugia to (1) modern standards and protocols for DNA collection and testing and (2) the release of the electronic data was the final blow for me in this extremely weak and flawed case.

With your permission, I want to leave a link to Michael Gaynor's latest article.

This raises key questions with regard to the "book" and its subject matter that Timothy Tyson has used to make a living for a lifetime, apparently.

This is a great article showing how someone can use an agenda all his/her life and will emerge to victimize others every time their "cause" inside the culture wars surfaces.

It has already been proven that Tyson removed pages from a thesis paper at an area library because they didn't square with his accounts in that little "book" that has been touted.

No doubt, at the time of that case long ago this country was going thorough enormous cultural upheaval; however, as everyone knows well, how Tyson and others at Duke and beyond were able to railroad innocent people is revealing enough to question all of Tyson's alleged "scholarship".

This also illustrates that in any criminal case, what takes place outside the courtroom can be just as significant to a case as inside the courtroom.

Very interesting what you have written and is easy for even me to understand. But as always, what I read causes me to ask more questions. And for anyone else, if you know the answers to my questions please jump in.

1. Does a strong reading of a full profile (high peaks or whatever) mean that DNA was deposited more recently than a low reading (low peaks or whatever)? Or is it always that a full profile can point to an individual but not the time frame of when it was deposited.

2. So if we have people who are "poor shedders" of DNA is it correct to say you can't determine innocence of an individual just because they didn't shed sufficient DNA at a crime scene in several places?

3. A lab has reference samples of those they are comparing DNA evidence against, correct? How do they obtain those reference samples from an individual (blood, saliva, hair, etc.)? And how are they collected and stored for comparison to crime scene evidence?

4. When you have DNA collected from a crime scene (especially the samples which contain more than one profile) can you tell what kind of DNA it is (meaning, blood, skin cells, saliva, etc.)?

5. How common is it in criminal/murder investigations to have a DNA mixture of two or more profiles?

6. I have read where DNA testing has helped free those who were convicted of rape (but were innocent) many years ago. Has this also been done in other areas of convictions (such as murder) where sexual assault is not present?

It is sometimes argued that Mr. Sollecito’s DNA could not have arrived on the clasp by contamination because his DNA was found on no other items except for a cigarette butt. This is essentially the reasoning of Judge Micheli, who oversaw Rudy Guede’s trial.

Another reason why this argument is flawed is that not even everything in Meredith's room was tested, never mind everything in the house. How do they know that nothing else contained Raffaele's DNA? And even when they did test an item, often only a couple of samples were taken from it (the bathmat, for instance). Just because nothing else they tested had Raffaele's DNA on it doesn't mean nothing in the house contained his DNA.

1. Does a strong reading of a full profile (high peaks or whatever) mean that DNA was deposited more recently than a low reading (low peaks or whatever)? Or is it always that a full profile can point to an individual but not the time frame of when it was deposited. I think that very little can be said about the time frame. If you have two samples that started out with the same amount of DNA and were stored under identical conditions except for length of time, then the sample that was stored longer would show a more pronounced decrease in peak height as the size of the DNA fragments got longer than the sample that was stored for a shorter period of time.

2. So if we have people who are "poor shedders" of DNA is it correct to say you can't determine innocence of an individual just because they didn't shed sufficient DNA at a crime scene in several places?I would say it depends on the circumstances of the alleged crime. It would be impossible for, say, three men to gang rape a woman over a thirty minute period and leave no DNA, even if they were poor shedders.

3. A lab has reference samples of those they are comparing DNA evidence against, correct? How do they obtain those reference samples from an individual (blood, saliva, hair, etc.)? And how are they collected and stored for comparison to crime scene evidence? One source of reference DNA is cheek swabs and another is pubic hair. I don’t know how the samples are stored.

4. When you have DNA collected from a crime scene (especially the samples which contain more than one profile) can you tell what kind of DNA it is (meaning, blood, skin cells, saliva, etc.)?Not to the best of my knowledge. To a very good approximation, the DNA in all of our cells is identical.

5. How common is it in criminal/murder investigations to have a DNA mixture of two or more profiles? I am not sure, but I have read that if you show a mixture to 10 experts, you may get 10 different interpretations.

6. I have read where DNA testing has helped free those who were convicted of rape (but were innocent) many years ago. Has this also been done in other areas of convictions (such as murder) where sexual assault is not present?I think that the answer is yes, but I cannot be sure. Prbobably the Project Innocence website would be a good place to check.

I just read the "DNA's Dirty Little Secret" article you linked on your last post. That was a bit of an eye opener.

I assume the electropherogram of a mixed sample would show all the alleles (genetic markers) of that sample together. It is then the task of the analyst to pick the alleles that match the known profiles.

Are alleles gender specific?

I ask that question because it has been reported that the bra clasp contained DNA profiles of at least four people - Sollecito, Meredith, and two females. I don't know if they were identified in the same sample or whether they were identified in different samples on the clasp.

It seems that working with a mixed profile of two individuals can be subjective. How reliable can a mixed sample containing markers from four people be?

It would be interesting to see how many markers from the tested sample matched Sollecito's profile.

I ask that question because it has been reported that the bra clasp contained DNA profiles of at least four people - Sollecito, Meredith, and two females. I don't know if they were identified in the same sample or whether they were identified in different samples on the clasp.

I think just one sample was taken from the clasp, and they swabbed both hooks at the same time. So they couldn't tell on which hook each person's DNA was found.

Apparently the reason they were able to identify Sollecito is because they used the extra-sensitive Y chromosome test to identify his DNA. His was the only male DNA on the clasp, while the rest of the DNA on the clasp was mixed female and thus trickier to identify.

Incidentally, Rudy's DNA was found on the main part of the bra next to the bra clasp, which originally led police to tell the media they thought Rudy was the one to cut the bra. I'm still trying to envisage a situation where Raffaele holds the hook of the bra clasp and Rudy holds the strap next to the clasp while one or other of them cuts it!

Mr. Moore comes to the same conclusion as Frank at PerugiaShock regarding the luminol enhanced footprints, the "clean up" and the source of the luminol glow, only with a searing indictment of Mr. Mignini and his forensics team.

Thanks; that is another excellent post from Steve Moore. I would like to add that according to Colonel Garofano in the book Darkness Descending, the luminol was not carefully applied in the hallway, so that the size of the footprint is hard to determine. Also, what Mr. Moore says about luminol and the color of the reaction being the same, no matter which catalyst is present, makes perfect sense. It is the oxidized form of luminol that is blue, and it is not important how the luminol got oxidized.

The prosecution alleged that the luminol-positive footprints were canceled bloody footprints. Those bloody footprints would have been made from Meredith's blood, yet Meredith's DNA was absent from all of them.

Amanda's DNA showed up in the luminol-positive footprints in her room. So Amanda cleaned up the bloody footprints and removed all traces of Meredith's DNA but left her own DNA in the prints in her room? I challenge anyone to explain how that could happen!

I'm sure someone has made a power point presentation to try to prove the footprints and the lack of DNA...

" Kate said...

Incidentally, Rudy's DNA was found on the main part of the bra next to the bra clasp, which originally led police to tell the media they thought Rudy was the one to cut the bra. I'm still trying to envisage a situation where Raffaele holds the hook of the bra clasp and Rudy holds the strap next to the clasp while one or other of them cuts it!

May 11, 2010 4:19 AM"

maybe Rudy was just trying to be a team player like on the basketball court? That sure is interesting positioning for Rudy's DNA and for RS DNA...the only possibility I can think of would be their hands would be there at different times.

In your opinion as far as the good vs bad shedders, it would be highly unlikely that only Rudy's DNA would be in great quantity in Meredith's room if there were three people involved in her murder and given the viciousness of the crime?

Randy wrote: maybe Rudy was just trying to be a team player like on the basketball court? That sure is interesting positioning for Rudy's DNA and for RS DNA...the only possibility I can think of would be their hands would be there at different times.

Ha, yeah, maybe Rudy was just being a good team player. No wonder the prosecution didn't try to argue Amanda's DNA was on the clasp as well. Imagine them trying to propose a scenario in which both Raffaele and Amanda were holding the hooks of the bra clasp as Rudy held the main part of the bra, and one of the three cut it... Almost sounds like Mignini's ritualistic sex game theory!

Here's the news article which mentions Rudy's DNA on the bra, from 16 December 2007, just before the bra clasp was collected:

Forensic experts in Rome have discovered Mr Guede's DNA on her blood-stained bra, which was torn off and left near her body. The traces are on the back of the bra, near the fasteners.

Thanks; that is another excellent post from Steve Moore. I would like to add that according to Colonel Garofano in the book Darkness Descending, the luminol was not carefully applied in the hallway, so that the size of the footprint is hard to determine. Also, what Mr. Moore says about luminol and the color of the reaction being the same, no matter which catalyst is present, makes perfect sense. It is the oxidized form of luminol that is blue, and it is not important how the luminol got oxidized.

Chris

This is the quote Mr. Moore uses in the article: "But Patrizia was not deterred by logic and fact. In court, she said that she knew that the luminol stain was blood “…because of its color.” Every trained forensic investigator (except, apparently Stefanoni) knows that luminol glows blue. Only blue. Not a different shade of blue for different substances…just blue. What Stefanoni said, she knew to be false, or she was so incompetent that she should not have been allowed to destroy a crime scene."

If I recall, we discussed previously several articles indicating that a trained investigator can in many cases tell if a luminol reaction is from blood or other substances. The reaction is stronger or brighter in the presence of blood, if I remember correctly. I don't know what the full quote from the above snipped version is. It is possible that she mis-spoke or it may be taken out of context or "lost in translation". If anyone has the entire quote, I would be interested in seeing it.

"How does luminol work?Luminol produces light during a chemical reaction with either molecular oxygen or a mixture of potassium ferricyanide and hydrogen peroxide and is probably the basis for most light sticks. In an alkaline (basic) solution, the luminol molecule becomes a dianion, a molecule with two negative charges on it. In this dianion form, the molecule has two nitrogen atoms exposed to the solution and these nitrogen atoms are easily replaced by two oxygen atoms. When that exchange takes place, a molecule of nitrogen gas is released and the final oxidized luminol is left in an electronically excited state. This molecule quickly gets rid of its excess energy by emitting light."

This guy comes off a bit like a smart-alecky teen; however, there's some interesting info there.

"Rose" is most accurate. Many shades emerge from its use.

Given the haphazard forensics milieu in the Knox/Sollecito case, I fail to see how any specimen of DNA can be deemed accurate or conclusive.

I found some quotes in Barni et al., Talanta 72 (2007) 896–913. This paper is linked at the Friends of Amanda web site, and I have discussed it previously in comments and one post.

“The most problematic chemicals for a correct interpretation of luminol test results are those which provoke intensification or a generation of a chemiluminescence emission even if blood is not present, leading to false-positive results. Due to the possible presence of these substances at the crime scene, the luminol test must not be considered sufficiently specific to permit an unequivocal identification of blood [15,18,51,88,89].”

“Of particular relevance in the forensic use of luminol are the interfering effects from plant peroxidases which are mostly abundant in fibrous plant material from fruits and vegetables (including the pulp and juice of fruits) but are also found with chemical variations among photosynthetic microorganisms [69,114]. In the presence of these enzymes, and to a lesser extent with other plant compounds such as the Mn2+ in Photosystem II [115] and/or the whole chloroplast [116], the luminol test can produce false positive indications often showing undetectable differences in light emission characteristics from that seen with blood [63,114].”

“Generally visual examination is used when the luminol test is employed in a forensic situation, rather than instrumental detection of the luminescence. An experienced practitioner maydistinguish the true blood-catalyzed chemiluminescence from that produced by other substances by the evaluation of parameters observable to the naked eye such as emission intensity,duration and spatial distribution. However this approach may also lead to misinterpretation, due to a subjective, informal and non-quantitative evaluation, for example, because its intensity isqualitatively much weaker than that expected for blood. In other circumstances an emission of similar intensity may be thought to derive from diluted bloodstains and is accepted. Therefore,caution should be exercised when using the test. Any confusion which may arise over a stain can usually be resolved by an intelligent observation and, if necessary, by further testing [73], for example, by using a different presumptive test for blood, such as the immunochromatographic test for the confirmation of human blood presence Hexagon OBTI (Human GmbH, Wiesbaden, Germany) [137].”

My own take on this question is that it is generally better to do a follow-up test. This avoids the subjectivity inherent in just using one’s visual interpretation. That goes double in this case, where the incompleteness of the footprint reference samples, incompleteness of the DNA reference samples, and other factors point to the forensics having been done in a nonobjective way.

Unequivocal innocence is an interesting concept but somewhat misplaced in the current context. Given that criminal trials are not undertaken to establish innocence--and certainly not 'unequivocal' innocence--but rather to establish the opposite. In this context, if DNA or other evidence is dubious, this tends to raise questions/reasonable doubts about guilt. Or, if you prefer, fails to establish unequivocable guilt.

So, it would be more correct to state that, 'given the haphazard forensics milieu in the Knox/Sollecito case, I fail to see how any specimen of DNA can be deemed accurate or conclusive.

"So, it would be more correct to state that, 'given the haphazard forensics milieu in the Knox/Sollecito case, I fail to see how any specimen of DNA can be deemed accurate or conclusive."*************************************

No, I said exactly what I wanted to say above.

You can sit in front of your computer and opine your way.

For all practical purposes, the ferocious Knox/Sollecito supporters---many of whom have displayed an uncommon rabidity on occasion---want to push a version of the idea of "unequivocal innocence".

I am reading Candace Dempsey's book “Murder in Italy,” and I would like to share one anecdote. After Raffaele Sollecito, Amanda Knox, and Patrick Lumumba were arrested, the police did not take them directly to prison. Instead they turned on their lights and sirens and drove through the old town in a kind of parade. The only time this had happened previously was when a mafia don was arrested. This incident and the photograph of Amanda on the wall of a police division in Rome before she was even formally charged speak volumes about the nature of the case. This case was not as big as the DL case; it was bigger, more like the Italian version of the OJ Simpson case. The perp walk in the DL case was ridiculous. The perp parade in this case was inexcusable, and it makes me wonder what the rules for a change of venue are in Italy. How could the defendants get a fair trial in Perugia after they had been put on display in this fashion?

The authorities released Mr. Lumumba about two weeks later, long after alibi witnesses and bar receipts had been available to show that Mr. Lumumba was elsewhere at the time. However, Mr. Sollecito and Ms. Knox did not have proof of innocence. It is possible that they never will be able to muster such proof, and some fraction of the reason for this lack is the way that the police mishandled multiple types of evidence. To my way of thinking, an equally important question is whether or not there was reasonable doubt of their guilt.

Dempsey's book sounds riveting and most certainly adds a significant layer of authenticity for the chemist in you.

Most informative that a spectacle was made during the arrests.

That aspect is quite troubling, indeed.

Just as a matter of course, I would be curious about the profiles, bios, and character of the Knox/Solliceto defense attorneys.

As we have seen in the Duke Lacrosse Hoax, an enormous charade of "heroism" and "fight for justice" will be the refrain from all defense attorneys.....

......when, in essence, they are just a part of the same system where prosecutors thrive and merely orchestrate a chess game for public consumption.

I cannot emphasize enough the degree to which the lacrosse defense attorneys have not been scrutinized for their longtime membership in the very "club" that gleefully tried to railroad Reade, Collin, and David.

At the time, Jason Trumpbour wrote eloquently criticizing and outlining the many bizarre ethical issues involved for lacrosse defense attorney James Cooney to have turned around and represented Duke University right after he pocketed the stash for representing Reade Seligmann.

The very act of doing such a thing tells me that Cooney is basically scum.....

.......yet no one wanted to highlight this odious scenario in which he participated.

Very bad for the "hero" schtick they had going.

In the future, any blogger who attempts to avoid discussing this aspect of the scenario should be taken as a mere instrument of the defense.

Perhaps engaging in mutual giddy and selective storytelling for the benefit of their own narrative.

If Liberal Democrats had not been "in charge" of everything during the Lacrosse Hoax, Crystal Mangum would have been put in jail for her high crimes.......

......and her subsequent arrest for attempting to murder someone would never have occurred.

Imagine those who play up her recent arrest.......

.......even as they did not push Cheshire, the "soldier" Bannon, and AG Roy Cooper to hold her accountable as you and I, or anyone else, would have been.

Personally, I have grown tired of these self-serving gymnastics.

Those of us who actually live in the Triangle and know the corruption that is alive and well inside the circles which the defense attorneys operate daily......

.......will not sit by and continue to see this fantasy played out without correction.

Wonder why the HBO film has not been made?

Could it be that the Triangle far left of which the lacrosse defense attorneys are a part does not want the real story told?

Because it would be revealing too much about the very milieu of which they are such a strategic part?

As I said before, it is an odious reality when you see the people who are their "buddies" and those whom they are now supporting politically.

That story needs to be told...and will be.

And in Italy........the Knox/Sollecito case could benefit from a totally impartial individual going over and spending months investigating the very heart and soul of that area and its "ministers of justice".

Defense attorneys assume their roles for grand compensation in the U. S.-----as we know so well.

It does appear to me that Steve Moore is not giving us the full scoop on the Luminol information just based on our own research. He is claiming that Stefanoni is either lying or incompetent. Yet the entire quote from her is “…because of its color.”.

I would not make a decision about her truthfulness or competence based this obviously out of context quote

Regardless of the shade/glow/promptness with which the glow began etc...aren't we inexplicably missing the DNA from this supposed blood? And without that where are we? Plus, my understanding is there was zero evidence (but much insinuation) offered at trial regarding the supposed clean-up...so that leaves us twisting in an onslaught of unsubstantiated claims that there was blood on the floor and that this blood somehow connected Ms. Knox and Mr. Sollecito with the murder.

"Unequivocal innocence is an interesting concept but somewhat misplaced in the current context. Given that criminal trials are not undertaken to establish innocence--and certainly not 'unequivocal' innocence--but rather to establish the opposite. In this context, if DNA or other evidence is dubious, this tends to raise questions/reasonable doubts about guilt. Or, if you prefer, fails to establish unequivocable guilt."

Absolutely brilliant ... that's a home run.

As you've noticed, there remain a few misguided individuals who insist that it is necessary for Knox and Sollecito to "prove their innocence" even adding a made up term, 'unequivocal innocence' a term that has absolutely no legal meaning whatsoever and is not even found in Black's, the Bible of legal terminology.

The proper focus as you correctly stated is "So, it would be more correct to state that, 'given the haphazard forensics milieu in the Knox/Sollecito case, I fail to see how any specimen of DNA can be deemed accurate or conclusive."

And that is precisely the essence of this case. This case is not and has never been about the 'Liberal Democrats' in the "Triangle" or the defense attorneys there. Such commentary is laughable in the extreme and could not be more irrelevant to this case.

Chris made an excellent point about highly prejudicial behavior toward and treatment of the accused, and both Observer and Joe made very thoughtful comments about the low quality of the evidence used to wrongfully convict the defendants.

And therein lies the injustice ... the prosecution never came close to proving guilt.

Has anyone here read the full Massei Motivations (and please, no Google translations, I believe one would go crazy trying to interpret 400+ pages of that translation method)? Not only is the report the jury's interpretation of the evidence presented at trial and reason for their decision of a guilty verdict towards Amanda and Raffaele I believe it contains references to the evidence and how it was argued by both the defense and prosecution.

I can't say there was or was not enough evidence to convict because I did not attend the trial. What I have read concerning the case does leave me with questions but I am reserving judgment until reading the entire motivations.

Also, has anyone read both Nadeau's and Dempsey's books? If so, where do they agree and disagree as to the case against Amanda and Raffaele?

Many are working on translations, including Amanda herself. I have read all of Ms. Nadeau's book and most of Ms. Dempsey's book. They are so different in the way that they approach the subject that it is difficult to compare them on a point-by-point basis. Ms. Dempsey's book is more in-depth and more insightful.

With respect to the footprints in the hall, I have a problem with the prosecution's assertion. If the footprints were set blood, then why were they not visible? If blood were cleaned (so as to make them invisible), then why don't we see streak marks showing a dispersion of the blood? That is my interpretation of what Steve Moore said.

Massei actually argues that the footprints were made in a residue of blood/water after the person had cleaned their feet, rather than being bloody footprints which were then cleaned up. Probably the reason he does so is that, as Chris suggests, if they were cleaned up there would have been signs of that. So the jury decided instead that these were invisible footprints which the person did not realize they were making.

Interestingly, this means there is no reason the footprints can't have been made in a similar way by someone treading on the bathmat and picking up the blood/water mixture on their feet, then treading it down the hallway. Amanda says she had a shower that morning and used the bathmat to dry her feet, of course. The judge's interpretation seems to open the way for the defence to make that argument.

(I learned that from reading the Google translation version though, so Christiana will no doubt disapprove! I actually like the 'rawness' of that sort of translation, and have had no difficulties in interpreting it. Any translation is only ever an interpretation of what was originally said, even the prettified, tidied up ones).

" Not only is the report the jury's interpretation of the evidence presented at trial and reason for their decision of a guilty verdict towards Amanda and Raffaele I believe it contains references to the evidence and how it was argued by both the defense and prosecution."*****************************************

Christiana has touched on one of the most significant issues of the case.

One suffered from contemporaneity.

Knox and Sollecito have been convicted.

Consequently, there is the burden of proving that those convictions should be set aside.

Proving "unequivocal innocence" would be a good way to do it as well as a path to a book and movie deal!

And you know Mom and Pop are busy behind the scenes on that avenue.

On a previous thread, I believe it was "Joe" who even grudgingly agreed with my point---the unavoidable reality that "innocence" must be proven now, for all practical purposes.

It would be important to learn more about the actual roles of the defense attorneys and their relationships with the prosecutor.

Most attorneys will say that they keep a "civil", "productive", and "non-adversarial" relationship with the prosecutor of a case.

How "non-adversarial"?

How "productive"?

How "civil"?

To the point where the client's best interests are jeopardized? To the point where the veracity of ALL facts are muted as a balm for ulterior concerns?

No doubt, more than a few people are ignorant of the fact that Black's Law Dictionary is not the Bible in the courtroom of public opinion......where most people are more concerned with what actually happened than what was determined in court.

A jury is asked to decide guilt.

"Not guilty" really means "not proven guilty".

It does not mean "innocent."

That's why NC AG Cooper's declaration of the lacrosse players' innocence was such a coup.

Now......as I have said before, it is important to have compassion for those who want to mulct from the Duke Lacrosse Hoax.

If only those aspects of it that appeal to their selective analogies.

Many have repeatedly referenced the lacrosse case as it pertains to prosecutorial misconduct, evidentiary issues, and the highly incendiary idea that young students have been railroaded.

All provocative and all provide an easy narrative.

But I would suggest that one become submerged inside the transliterate conventions of the Italian language, the Italian culture, and the extremely complex relationships between any legal defense team and the prosecutor of a case.

To comment intelligently and definitively on this matter would require one to take up residence---not just for a few weeks but for months or years!---in Perugia and actually learn the dynamics of their court system and the community.

How well did the defense attorneys perform for Knox and Sollecito?

Do they know the prosecutor well? Do they like him?

Do they fear him?

Were they similar to some lacrosse defense attorneys in wanting to play the game for public consumption that the prosecutor was acting in good faith.....

.......letting things slide as they pretend not to know that the guy is/was dirty......

(I learned that from reading the Google translation version though, so Christiana will no doubt disapprove! I actually like the 'rawness' of that sort of translation, and have had no difficulties in interpreting it. Any translation is only ever an interpretation of what was originally said, even the prettified, tidied up ones).

Kate I don't disapprove of you having the patience to read a Google translation of the Massei Motivations. I wish I had your patience. I'm sure when sifting out the errors one can get an overview of the content of the translation, however, I think some accuracy of content may be lost.

Reading the motivations in Italian is the best course, second best would be to read it in a responsible translation in one's own language.

If I had a choice I would rather read the court transcripts then read the motivations.

Chris,

Maybe you can answer this concerning the two books: do they confront the case from a prosecution or defense viewpoint or from a neutral reporter's viewpoint?

Barbie's book is written from a pro-prosecution point of view (the same is true of Darkness Descending), and Candace's book is implicitly pro-defense. However, Candace is much more analytical, and she picks apart logical flaws in the prosecution's arguments that have not always been explored elsewhere. I think that Candace is more objective than Barbie is. She is certainly more thorough.

I think it would be good, then, to read both. It is always good to hear/read/research both sides of a subject.

Debrah,

You raise some good questions.

I'm not so sure that Amanda and Raffaele have to prove their innocence to the court. If on appeal they can have some or most of the evidence against them thrown out I would think their convictions would be overturned. On a public level, I do agree that they have to show they could not and did not kill Meredith.

Christiana,Good advice on reading both. I plan on reading Barbie's first just because I think she is a better writer than Candace from what I have read of their articles. It is no surprise that Chris is going with the more analytical one.

Based on Chris's posts about independent reviews of DNA evidence and the need for the raw data (.fsa files), it seems one way Knox and Sollecito could prove their innocence is if the appeals court compels the prosecution to turn over this data and allowing/ask for an independent review.

It's apparent that the prosecution has no interest in voluntarily turning this over.

The defenses didn't even know the machine read "too low" on The Knife DNA or the quantity of DNA on the bra clasp until a year and a half after Knox and Sollecito were arrested. This only occurred after Massei compelled them to turn over more info during the trial, in June or July of 2009.

A real coupe would be reconstructing the activities on the accuseds' computers from the night of the murder. It isn't clear whether Amanda's computer hard drive has been turned over to allow the manufacturer to salvage whatever can be salvaged of it's memory.

Unbelievable that. How on earth did the police manage to fry three hard drives? If that doesn't make you wonder about prosecutorial misconduct, then nothing will.

I think about the only thing AK's computer would show is some details on her and MK's relationship perhaps...I've always thought it kind of odd that the hard drives couldn't be salvaged. I wonder how much effort was actually put in to that.

Secondary DNA transfer...perhaps this can explain RS DNA on the clasp. maybe Amanda grabbed the bra after contacting RS or MK touched the towel RS used. Didn't MK and AK share the same bathroom. I have no idea how much DNA would get transferred in this way. I think MK's DNA on the knife can come from the lab like in the salmonella reference in Chris' article. Someone touched an area that was not cleaned and then it was transferred to the knife. Especially since the blood test didn't show the presence of blood.

The article about luminol on the injustice site was interesting since it mentioned about how streaking would appear in cleaned up areas.

I can see skipping the obvious bloody footprints for testing for blood. After all, you have a bloody murder and footprints in blood leading from the body to the exit door. I believe when Stefanoni was talking about the color in the above quote, she may have been referring to the color of the bloodstain itself rather than the color of the luminol reaction. Again, it is hard to tell with a snipped, out of context quote from some document or testimony that is not available to us peons. Steve has not decided to "share" this entire quote with us, in context.

On these obvious visible footprints, I see it as understandable that they would be more interested in the DNA results because it was simply common sense that the bloodstains were blood.

Six weeks later was when the luminol testing was done for footprints that were not visible. I see it as very possible that Mr. Moore is referring to these prints in his critique and Ms. Stefanoni is referring to the obvious bloody prints in her quote.

Amanda's computer would have had photographs of her and Meredith, thereby helping to take the air out of the prosecution's claim that the two did not get along. The last I heard, the defense offered to pay for Toshiba to look at it, but they were refused.

The loss of the information of when the two watched Stardust goes more to issue of an alibi.

I think you are not taking into account Stefanoni's other deviations from sound science, such as her assertion that skin cells do not contain DNA and that the placement of DNA on the handle tells us how Amanda used the knife. These chip away at her credibility before we even get to the manifold shortcomings of the knife profile itself or the bra clasp. Mr. Moore makes a good case for doing it the right way, IMHO. I also do not agree with you about what Stefanoni meant by the color. I think she was referring to the luminol blobs, but you might have to page through a bunch of posts at JREF to find it.

The blob you linked in another post looks like a Rorschach test. I don't think it looks like anything, and I don't see anything that looks like streak marks. Even Colonel Garofano implied as much in Darkness Descending. In addition if there were a cleanup in the hall, why didn't they clean up the bloody shoeprints? Sorry, but I do not think there ever was a cleanup.

Joe,8 bloody shoe-prints were found in Meredith's room, 5 of which were on the pillow under her body. The other 3 are close together at the foot of Meredith's bed. The lack of other shoe-prints is odd. It would be hard to mop the pillow and they could have missed the spot at the end of the bed. There were no footprints (as opposed to shoe-prints) found in Meredith's room, that I recall.

It is a mystery to me how this is possible without some sort of cleanup even if Rudy was acting alone and Amanda and Raffaele never went in to that room.

That is possible Chris. My theory is that he sat down on the end of the bed and removed his bloody shoes before doing that possibly with a side trip to the bathroom to cleanup, then back to the bedroom to put his shoes back on, leaving that trail exiting the flat.

Rudy's DNA was on the bra but not the clasp. If Raffaele lifted the bra up by the clasp as Amanda cut the bra with the "double DNA knife" it could explain this, even if (as is my opinion) the knife was not used in the murder. That minute amount of Meredith's DNA could have been transferred from the bra to the slight indention in the knife and Raffaele's DNA could have been transferred to the clasp.

That would put Amanda and Raffaele taking part in the cover up (staging the murder scene) but not necessarily in the assault.

I do not put any stock into the argument that there is too much of Raffaele’s DNA on the clasp for it to be there by anything but primary transfer. This line of reasoning runs counter to the consensus view that a DNA profile itself rarely provides us with clues as to how it was deposited. There is certainly enough DNA in fingerprints for a DNA profile to be developed, and Raffaele’s fingerprints were on Meredith’s door, IIRC. Towels are known to pick up DNA, and one unpublished study has indicated that they can be involved in tertiary DNA transfer. Also, a secondary transfer or contamination event need not take place in Meredith’s room; there are a couple of routes for DNA transfer to occur in the lab. For example, a technician could handle the cigarette butt then the clasp. Or equipment that touches one evidence item is then used on a second item, as is documented in this post. Finally, I do not know whether or not the police ran or reran Raffaele’s reference profile at the same time they ran the clasp, and the reference sample is a potential source of contamination also.

When the polymerase chain reaction is used to amplify the gene for amylogenin, two different sized DNA pieces are generated from the X versus the Y chromosome. This is how the gender of the DNA donor is often established. However (to answer your question), the alleles on the other chromosomes are not gender-specific. There is also a special type of DNA forensic analysis called Y-STR (STR stands for short tandem repeat) that is only applicable to males, and this test was consistent with Raffaele’s DNA profile.

an earlier post mentioned Rudy's DNA being found on the strap next to the clasp.

I came across some photos Frank posted on his Thursday, October 16, 2008 Pointing at the Murderer II article...http://perugia-shock.blogspot.com/2008/10/pointing-at-murderer-ii.html

(Frank says they are luminol but I believe they are actually from something similar- http://www.bluestar-forensic.com/index.php)shows an example of a cleaned table- http://1.bp.blogspot.com/_mqKZSZYw_HQ/SPgiIHYSKeI/AAAAAAAAAxI/15TS5ARF1cA/s1600-h/cleaned+table.jpg

this one is a cleaned floor-http://4.bp.blogspot.com/_mqKZSZYw_HQ/SPgiBK2N_iI/AAAAAAAAAxA/gFIrNSgTavc/s1600-h/washed+floor.jpg

"For what reason would Amanda and Raffaele take part in the staging of the crime scene if they didn't have a part in the assault? Is there any theory where that scenario would make sense?

I am not being harsh with you - I am wondering if you really think that is possible and why?

Christiana"

I believe the evidence seems to show they had some involvement in the staging of the break-in and the murder scene. That question of motive here is a tough one, but not as far fetched as the motives given for the murder (if they were involved).

It is possible the staging was done to cover a subsequent crime (theft). I wonder if something like a stash of drugs was taken from Meredith's room. Same is possible for Filomena's room. Would she have reported that?

The events of the morning of the discovery of the break in and letting others "discover" the body seems almost scripted.

I believe the evidence seems to show they had some involvement in the staging of the break-in and the murder scene. That question of motive here is a tough one, but not as far fetched as the motives given for the murder (if they were involved).

Rose,

Interesting theory.

I find it difficult to believe that Amanda and Raffaele would come back to the crime scene to stage anything if they weren't involved with the murder. It would be too traumatic, not to mention too risky.

I'm beginning to understand better some of the science behind DNA transfer and contamination (though "tertiary" is a new term I will have to look up). Thank you.

Here are two articles I found interesting. They may already have been discussed in prior posts but I missed the discussion. One article is from a few months ago, the second is from last week which is not DNA related but deals with human errors and/or negligence in crime labs (sorry, I forgot the lesson on how to do direct links).

In my May 13, 2010 6:19 PM post, I referred to the use of sirens in the parade on November 6, 2007. I should have said honked their horns instead of turned on their sirens. They also turned on their headlights. I am sorry for this error, but I still have the same problem with this action as before.

Tertiary transfer takes place when person donates DNA to something which donates it to something else which transfers it to a final object. tertiary transfer is like secondary transfer, but there is one extra step.

"... The events of the morning of the discovery of the break in and letting others "discover" the body seems almost scripted. ..."

It can be argued that there is no clear picture of what the actual events of that morning were, without the various embellishments given in media reports. That aside, the above asserts that the accused wanted others to discover the body. A simpler explanation is that they did not wish to find a body. I doubt many of us are keen to discover a corpse in a room in our homes. The only thing that would motivate me under the circumstances given would be the sense that someone needed my help. Short of that, I would beg the police to do the breaking down of the door for me.

I agree that Amanda and Raffaele wanted others to discover the body. They told of how they banged on that door and Raffaele attempted to break it down. As soon as he "made a crack in it", he stopped trying. I don't know about you but making a crack in the door shows progress towards the goal of opening that door. Stopping at that point does not make sense.

One of the young lads downstairs made a few kicks on it and got it open. The break in that never happened was designed to point suspicion away from Amanda should Filomena tell the cops that her stash was taken. The arranging of the murder scene to look more like a sexual assault was also designed to point the finger away from Amanda. Amanda's lamp in the room was to give them light to search for Meredith's stash and to arrange the body. The phony calls to Meredith's phone, the call to Raffaele's sister and calls to Filomena were designed to give credence to the scenes. Stepping away from Meredith's door at the time it was broken down was probably a mistake. Amanda had already seen enough and did not want to see it again. Raffaele was broke and Amanda had just paid her rent.

It is just a theory. I don't think the evidence is solid that Amanda killed Meredith yet I do think it is pretty clear that Rudy did not enter by Filomena's window and that window was broken from the inside. It is also clear to me that Raffaele and Amanda are hiding something. If there is a better theory that would explain the possibility of Amanda taking part in the staging but not the murder, I would love to hear it.

"...It is also clear to me that Raffaele and Amanda are hiding something. If there is a better theory that would explain the possibility of Amanda taking part in the staging but not the murder, I would love to hear it. "

Do you mean that, because you believe that Amanda participated in the staging, we must conclude that she also participated in the murder? I think that more than 'belief' is required to accept the conclusion that there was a staging in the first place. The evidence just isn't that clear.

In the case cited, I wonder if the second lab which tested the re-enactment of the tertiary transfer tried to force the results of the profile/s same as the first lab?

I guess tertiary transfer is possible but I would rather argue contamination or secondary transfer in my favor (rather than tertiary transfer) should I ever have a case brought against me.

As to your correction concerning Patrick's, Amanda's and Raffaele's arrest: Police departments would serve themselves better to drop the theatrics and issue a statement saying they have a suspect or suspects in custody, no more, no less.

Rose,

I haven't any theory but I can't wrap my mind around someone having no part in the murder and yet staging the crime scene to cover a theft. Better to be convicted of stealing than to be convicted of murder (or a passive accessory to murder).

Rose wrote: Rudy's DNA was on the bra but not the clasp. If Raffaele lifted the bra up by the clasp as Amanda cut the bra with the "double DNA knife" it could explain this, even if (as is my opinion) the knife was not used in the murder.

I don't really understand the police claim in Frank's article that Raffaele's DNA was on the 'perfect place' to hold if you were cutting the clasp. His DNA was on one of the *hooks* of the clasp, not on the fabric of the clasp itself, which would be covered if someone were wearing the bra! On the other hand, Rudy's DNA being found on the strap 'adjacent' to the clasp makes a lot more sense.

I agree that Amanda and Raffaele wanted others to discover the body. They told of how they banged on that door and Raffaele attempted to break it down. As soon as he "made a crack in it", he stopped trying. I don't know about you but making a crack in the door shows progress towards the goal of opening that door. Stopping at that point does not make sense.

It makes sense if he was ambivalent about breaking the door down, and didn't want to do damage to property with no good reason. Maybe he stopped *because* the door cracked, and he didn't want to take responsibility for damaging it further. After all, the police didn't want to break it down either, and let Luca do it. That's the problem with so much of this behavioural evidence: there are equally plausible innocent explanations for all of it.

Examine anyone's behaviour in minute detail and there are going to be oddities and inconsistencies - it would be easy to create suspicion about Filomena's behaviour that morning, for instance. So worried about Meredith, yet didn't bother calling her. Managed to call Amanda four times though. Did she already know Meredith wouldn't answer? Supposedly found the blood in the bathroom incredibly alarming, yet didn't bother heading home herself till Amanda told her about the break-in. Very insistent that the door be broken down - did she know what was in there, and make sure she was near the entrance to explain any traces of herself that might be found? Said Amanda called her at 12:35, when it was really 12:08 - why? Just silly examples, obviously, but it would be very easy to create a narrative of suspicion from her behaviour that morning.

I have a real problem with the way the judge relies so much on this very subjective interpretation of behaviour. The only way this sort of thing becomes significant is if you already *know* someone is guilty, and look to their behaviour to explain things in hindsight. It's no good as primary evidence of guilt, because there are too many possibly innocent explanations for it. This is another area in which the judge begins with the *presumption* of guilt, and examines the evidence from that standpoint.

Rose said: The arranging of the murder scene to look more like a sexual assault was also designed to point the finger away from Amanda.

The judge/jury appear to have rejected this staging of the murder scene, though. We don't know what was said during the closed sessions, but apparently it wasn't enough to convince the jury anyone went back into the room to stage anything. They thought the clasp was cut during the assault.

Amanda's lamp in the room was to give them light to search for Meredith's stash and to arrange the body.

I'm a bit suspicious about the lamp - if you look at photos of the crime scene, the plug and cable of the lamp are leading out into the corridor. Yet it can't have been there when the door was broken down; one of the eight people present at the time would surely have mentioned it. That means one of the police officers must have picked up the plug and lamp cable and placed it in the corridor. Why?

Or is it possible the lamp was used by one of the first officers on the scene, and became part of the crime scene inadvertently? Only the police would have a motive not to use the two lamps (desk/ceiling) already in the room. (Battistelli was in there on his own at one point, explaining what he could see to the carabinieri on the phone; if there's an unreliable witness, it's him). That would certainly explain the prosecution's reluctance to introduce it as evidence.

I also think that if Amanda and Raffaele had left it there, they would have mentioned it to the police along with all the other odd things in the cottage. They had absolutely nothing to gain by not talking about it.

Joe,The funny thing about this theory is that those on the guilty side of things seem to hate it almost as much as those on the side of innocence. Christiana can't wrap her mind around it and I can't wrap my mind around Amanda and Raffaele participating in the murder. However, the evidence seems to me to be convincing that they participated in a cover up and I remain unconvinced that they participated in a murder. This theory is just my feeble attempt to reconcile those two positions. I recognize that.

I just finished reading Angel Face by Barbie Nadeau. Most of you are aware that the "guilters" seem to think it is the definitive work on the case thus far and the "innocents" view it as a load of rubbish. It is a short book, a little over 200 pages and could have easily been printed on as little as 150 pages or so. It's an easy read but if you are looking for any great revelations, you will probably be disappointed.

As I read it I had my post it note pad handy and marked those pages that I found to be interesting, odd, or unusual. I will comment on these things first. As in my theory above you might find what I decided to focus on a bit strange. Try to overlook that and pay no attention to the girl behind the curtain.

Nadeau's comments about Frank at Perugia Shock were very interesting. "He was the quintessential blogger-a smart, cryptic insomniac. Even the chief prosecutor, Giuliano Mignini, read his posts." I get the impression that the press (Nadeau included) were somewhat jealous of Frank. He seemed to "curiously" have documents and information that the rest of the press did not have or didn't get until after Frank got it. The rest of the press seemed to meet at bars after court and discuss the case on a fairly regular basis. They had a system of barter or exchange where they would trade information and documents that other reporters did not have. I see evidence that this is still going on today in the press, as well as on the blogs dealing with this case. It is unfortunate, in my opinion, that sources do not reveal themselves or information is traded with understanding that the sources are not to be revealed in exchange for the information. In some cases, these tidbits of information or documents are held on to and closely guarded as assets in this barter system. It seems to be a case of I have this and you don't kind of thing and who knows if the information is truth or falsehood?

According to the book, Mgnini believed that Frank's blog was funded by Mario Spezi (see Monster of Florence and Douglas Preston references). According to Nadeau, Mignini allowed her and Andrea Vogt to listen to a wiretapped conversation between Spezi and Preston and in Nadeau's opinion "they do seem to refer to steering the police to faked evidence".

I get the feeling that Ms. Nadeau believes Amanda and Rafaelle would have been found innocent of murder with a more effective defense. Nadeau describes this defense as disjointed and in some cases the teams of Knox and Sollecito seemed to work against each other. I think the best parts of this book deal with the descriptions of the lawyers and prosecutors as well as some of the characters and expert witnesses. There is one passage about a forensic expert named Vincenzo Pascali on Raffaele's defense team that quit over DNA findings that would have been harmful to the defense. According to this book, Pascali believed that there was evidence of Amanda's DNA on the bra clasp as well as Raffaele's.

My overall impression of Angel Face is that it was written by someone used to writing brief articles and compacting a lot of information into just a few words. The book is missing a lot of detail. No documents, no photo's, no forensic articles, hardly any quotes on actual testimony, and little explaining of the writer's opinions that are given during the course of the book. In short the book falls well short of a definitive designation. I believe the drug culture in Perugia is over-stated and some of the information (like the bleach receipt) is not correct.

Her descriptions of witness testimony seem to favor the side of the "guilters". Nadeau has little to say about the believability of some of the witnesses for the prosecution, yet she describes many such problems with the defenses witnesses. Her slap down of the defense ballistics expert and how he "slunk away" after his testimony is a classic example, in my opinion.

It is also my opinion that Nadeau makes a pretense at being objective. It is clear to me she favors the side of guilt yet does want to appear to be open minded. The book has a difficult chronology, skipping in time and subject material and appears to be poorly edited, in my opinion. I get the feeling it was rushed to press at least one or two re-writes early.

I will read Candace Dempsey's book next and I hope it is better than this one.

Your post is helpful and leads one to believe that the charge of 'face saving' is more applicable to members of the press than to any of the parties directly involved in the case!

Question: you say that "[a]ccording to this book, Pascali believed that there was evidence of Amanda's DNA on the bra clasp as well as Raffaele's." Does Nadeau attribute this to Pascali directly, or does she just imply it?

To Anon at 9:22AM :Thank you for that kind comment. Here is the actual passage dealing with that subject:

"He had been hired to discredit the bra-clasp evidence. But when he started hinting that in his own findings, the sample also contained Knox's DNA, Bingiorno objected. Introducing Amanda's DNA on the specimen would only make things worse, tying both defendants to the crime scene."

" ... Why is it that so many want to avoid discussing the motivations, or lack thereof, of defense attorneys in such cases?"

When 'motivation' is mentioned in relation to the Italian case, it refers to a legal document that judges are required to prepare after a verdict is rendered to explain the reasoning behind the verdict. There's no similar requirement for any of the attorneys. If you intended to start a discussion of what motivates attorneys, how does that relate to the strength of DNA evidence or likelihood of contamination?

"If you intended to start a discussion of what motivates attorneys, how does that relate to the strength of DNA evidence or likelihood of contamination?"*************************************

"Anonymous"--

If you "intended" through your delicate snide remark to shut down discussion of all else but the precious DNA, which, in this case, seems to have been handled so ineptly, please just spell out YOUR "motivations".

But if you ever become so daring as to post using anything but anonymity, please try hard to comprehend that use of the word "motivations" was intended for a generic version.

Not the legal document used by the judges.

But you already knew that, didn't you?

When you choose to gloss over the "motivations" of anyone in any scenario of life---criminal trial or no criminal trial---you will be losing out on most of life's meaning.

Regarding the motivation, or lack thereof, of defense attorneys in such cases, I too have wondered that.

My current impression is that they did not have a chance of winning this first trial. The DNA and luminal stuff goes over my head but I can generally follow the staged break-in theory. If you take a real close read of this section

http://forums.randi.org/showpost.php?p=5834900&postcount=7036

of the Massei report and take note of what is accepted/rejected as evidence and the underlying logic, it will give you a flavor the mindset of the jury and what the defense was up against. I really don't think they stood a chance at this stage.

I would be grateful if we could all keep snide comments and sarcasm to a minimum.

To all,

Darkness Descending implies (correctly or not) that as far along as the defense summations, there was doubt among close observers about whether or not Sollecito's defense team (led by a high profile political figure) would try to separate RS from AK. The opposite actually happened in the close.

Thanks Chris, for your heading off what we all know will be attempted without your close monitoring.

Since I already know with whom we are dealing just from my own Site Meter, I would encourage your vigilance on what continues to be someone from afar attempting to create discord for no apparent reason.

An interesting side note to that translated portion of the motivations report dealing with the staged break in. Nadeau relates how they asked the defense expert if he was able to hit his target the first time and he admitted he was not able to and had several misses. I imagine that big rock would make some kind of mark or gouge on the wall if you missed the window.

As Chris related previously they then got into a discussion as to why he threw the rock over-handed rather than under-handed. LOL.

Welcome Humanity Blues.

Kate, An interesting theory on the lamp. I for one like to hear other theories on what may have occurred.

I've had my blog for about 3 years actually. The basketball blog I had I was just too lazy to start, and my other blog I'm reformatting, so it is locked for the time being.

I meant "subscribe" by email btw, which means you have to post something before follow-up comments will be sent. See now look what you made me do? Now I'm all on the offensive and I didn't even do anything!

Anyways Chris, keep up the good work. You take enough piss at JREF and I think you have a very good blog.

Thank you for the review of Nadeau's book. I was also given a summary of the book by Humanity Blues last night. Maybe if you both write a bit more I won't have to read the book myself. I do hope to fit it into my schedule soon. Then Dempsey's book and Darkness Descending.

And Rose, I wasn't criticizing your theory. It doesn't take much for me not to wrap my head around something. Your theory is as valid as the one Massei and jury came up with as regards to motive.

I think the defense attorneys motivation in this is case would be to get their client off in one of the biggest and certainly one of the most publicized trials in Italy. I thought I read that one of AK's lawyers was going against Mignini in 3 or 4 other cases. They end with a not guilty verdict for AK or RS and they are famous and probably would get a few more euros for their work.

I wonder how much time RS superstar lawyer- Giulia Bongiorno spent on the defense. If she was the only lawyer for RS, would his outcome had been different? And how good were AK's lawyers? I thought I read that one of them wasn't very experienced in this type of case so they hired a 2nd lawyer.

Going back to the hard drives that were messed up by the lab.

http://www.deadharddrive.com/

This is someone who repaired his own HDD after replacing the little logic board. It doesn't sound too difficult to me. I don't think there was any benefit to the police to get them properly repaired so they were never fixed.

http://www.bluestar-forensic.com/gb/bluestar-bleach.php

At the bottom of the page there is a video- about a 5 minute video excerpt showing bleach and blood reactions with this bluestar product...mostly shows bleach reactions.

Yes, I am 3:46 p.m. I try hard to remember to sign my posts but do forget from time to time.

Thanks for the link. I have not read any excerpts from Dempsey's book but will do so on your recommendation. Chris also linked an article on JREF I am reading. Science stuff but it's interesting. Read it if you get a chance.

Analyis of DNA mixtures is complex, and from what I can gather, has elements of subjectivity. If independent experts could get copies of those pesky electronic data files, we might be able to obtain a clearer picture of whose DNA might be there.

You are probably right about those pesky whatchamacallits, Chris. Once they get them it should be a simple matter to show the tertiary transfer of an octogonal contamination mixture caused by malicious and diabolical dirt devils.

Or it could be the defense expert was an honest man and also better at reading this DNA stuff than the prosecution experts and was forced to quit so he would not reveal the truth.

Of course, after he stuck Raffaele with a bill for 50,000 euro's he probably felt somewhat better about the whole sordid mess.

Analyis of DNA mixtures is complex, and from what I can gather, has elements of subjectivity. If independent experts could get copies of those pesky electronic data files, we might be able to obtain a clearer picture of whose DNA might be there.

Will one profile in the mixture always be of higher volume than another? Are female profiles higher volume than males in a mixture or the other way? If the mixture is a result of a crime wouldn't both profiles be of high volume?

I don't think that there is any systematic difference between males and females. I don't see any reason why two individuals could not contribute equally to an item of evidence.

Rose,

One wonders why the prosecution did not argue that Amanda's DNA was on the clasp.

A pro-guilt crowd's argument about the bra clasp is that the two of the unidentified females who contributed are Laura and Filomena, and their profiles are present due to shared laundry facilities. I have criticized that argument on the basis that we do not have Laura's or Filomena's reference profiles. But suppose that they are right, and two of the profiles are Laura's a Filomena's. Then why can't the third profile be Amanda's, deposited by shared laundry facilities? The presence of DNA almost always tells us nothing about how or when it was deposited.

If profiles in a mixture are uneven in their volume (intensity?) it would have me wondering if the lesser profile/s weren't already deposited sometime prior to a crime being done. Of course, I can have this all wrong.

I think the obvious reason as to why the prosecution didn't argue that Amanda's DNA was on the clasp as well is that it would have greatly weakened their case if they'd tried to do so. What theory could they possibly have come up with to explain how Amanda and Raffaele (and a couple of other people) left their DNA on one of the hooks of the bra clasp and nowhere else on Meredith's clothes, body or possessions? What bizarre scenario as to how the bra clasp was cut could possibly have explained Rudy's DNA next to the clasp, and Amanda and Raffaele's on one of the hooks?

If the prosecution had tried to argue Amanda's DNA was on the clasp it wouldn't have made the case for her and Raffaele participating in the murder stronger, it would have made it weaker and drawn attention to the likelihood of contamination. So they only argued Raffaele's was there, because they needed evidence against him (this was just after the shoe prints had been proven to be Guede's).

Something I'm still a bit confused about is the amount of each person's DNA on the clasp. I know that the prosecution and defence experts were arguing in court as to the ratio of Meredith to Raffaele's DNA on the clasp. The defence expert argued the ratio was 10:1 and then 8:1 when he was given more information, which puts the amount of Raffaele'e DNA within LCN range (i.e. below 200 RFU). Stefanoni said it was 6:1, and therefore not LCN. A figure of 1400 RFU has been mentioned I think, and this sort of makes sense with the ratios mentioned above - 8:1 giving a figure of 175 RFU, and 6:1 giving 233 RFU.

So that would seem to be pretty low. But is there another way of measuring it, i.e. in terms of the amount/volume? And what were the figures for the DNA from other people found on the clasp? Can they even tell the amount with any accuracy, given that it's a mixed sample?

Kate,That is an interesting possibility regarding Meredith's DNA on the bra clasp or not. My main interest is to determine if it is true that her DNA was there. If the review of the DNA evidence that the defense has asked for in the appeal is granted, maybe the results of this will be reviewed again.

If the prosecution had tried to argue Amanda's DNA was on the clasp it wouldn't have made the case for her and Raffaele participating in the murder stronger, it would have made it weaker and drawn attention to the likelihood of contamination. So they only argued Raffaele's was there, because they needed evidence against him (this was just after the shoe prints had been proven to be Guede's).

Kate,

There may not have been what is considered a full profile of Amanda's retrieved from the bra clasp for the prosecution to argue anything. There may have been some markers compatible with Amanda's profile but not enough to argue it was hers. I don't know what is considered a full profile so this is just speculation on my part.

I prefer to define the LCN region on the basis of when stochastic effects start becoming noticeable or on the basis of how much DNA template is available. The problems with defining it on the basis of RFUs start with the fact that the signal strength is dependent upon many things, not the least of which is the number of cycles of the polymerase chain reaction that are employed, and that is under the control of the experimenter.

The issues with respect to how much DNA are on the clasp are complex, and I do not think that there is enough information available publicly to answer them. It was the question of how Stefanoni had determined this number but had not conveyed it to the defense that precipitated the defense asking the court to compel complete release of the DNA forensic information last summer.

Christiana, yeah I think you might be right about Amanda's profile on the clasp only being partial (I'm not sure but I think that may have been mentioned on Frank's blog).

I would still argue, though, that if it had benefited the prosecution to try and argue the DNA was Amanda's, they would have done so. Even if there'd been, say, only a 70% chance the DNA was hers (which I'd think would be quite low) surely the prosecution would just have thrown it in there as an additional bit of damning information? After all, the dubious testing on the knife DNA makes that piece of evidence far less reliable than the profiles on the clasp, which were at least obtained using conventional, approved testing.

Stefanoni could just have said, 'a partial profile for Amanda also appears on the clasp', and let the jury make of it what they chose. The fact she didn't do that makes me think that arguing that point would have damaged the prosecution case more than benefited it. (Well, I think it certainly *would* have damaged the prosecution case more than benefited it, whether or not that was the reason behind Stefanoni choosing not to make that argument).

Thanks for that information on LCN, Chris. I've heard it mentioned a lot that the amount of Raffaele's DNA on the clasp was greater than that of the other DNA contributors, but there usually aren't figures to back it up so I was wondering if there was some definitive answer. Seems not! It's all a bit vague.

Rose, thanks for your link to the report - I just noticed you also have Mignini's translated sentencing report up there too. Excellent, more light reading. :D

Thanks Kate.Ray Turner has a new post up about Mignini and he has even decided to have comments allowed.There is a bit of trouble posting those comments as of this moment but I am sure he is working on it.

I don't agree with Ray on much, but I think he is one of the good guys. I will give him my unofficial snark post whenever he fixes the problem. LOL.

Comments are working at Ray Turner's place now. I decided not to post a snarky reply and instead went with my gut feeling that Mignini is not quite right in the head. It has taken awhile for me to come to this conclusion and I believe it fills in some of the questions I have about the strange series of unfortunate happenings to Raffaele and that Knox girl.

The problem I have been having is trying to fit what I believe the evidence shows to what actually happened the night of the murder and the following morning. I am no longer convinced that the evidence is contradictory simply because some of the evidence may not be true evidence, in my opinion.

Kate said... Rose, thanks for your link to the report - I just noticed you also have Mignini's translated sentencing report up there too. Excellent, more light reading. :D

Thanks, Kate.For any interested the complete Italian version of Amanda's Motivation report is there as well. And I have recently added a Google translated version of Rudy's motivation report from the December appeal decision (he has another appeal underway now). Amanda's and Rudy's translated reports read much better than the translated Mignini report for some reason. I read thru that translation twice yesterday and came away needing serious amounts of alcohol.

I read thru that translation twice yesterday and came away needing serious amounts of alcohol.

Be careful Rose. Mention of your alcohol intake has put your posts into Abandon All Hope at JREF.

I do want to thank you for all the work you have done converting pdf files into word documents. Also, thank you for reading the translated English versions of the various motivations. I haven't done so - it is easier reading the Italian. :)

And Chris, I hope all is well and you will be back posting on a regular basis soon.

According to Frank, Mignini contends that Amanda slandered thirteen officers and the interpreter. So, there were thirteen officers involved in her interrogation?

According to Perugian authorities, Amanda was treated well and freely (without coercion) accused Patrick of the murder. If she so freely spilled the beans as they contend, why were thirteen officers involved in this interrogation? That would be completely unnecessary.

The only reason so many officers would take part in or be instructed to take part in an interrogation would be to intimidate, confuse, and coerce a suspect.

Thanks Joe,I never thought of this bit about 13 officers involved in the interrogation. This is more than overkill. A young girl facing thirteen people in a foreign county would be terrified. Is it no wonder that she can't identify the person who slapped her. TM

This is interesting that JOE and others commentators assume: "As argued by you and others, the prosecution should turn over all data, including the electronic data files, to the defenses. I don't see how a court of law in a civilized nation, whether it be Italy, USA, etc., could justify such a refusal."

But none of you knows or care to learn that Italian criminal law offers the defense to be present during the forensic examination ( of the knife in this instance). Except that defense lawyers of Knox and Sollecito declined the offer.. .Speaking about civilized countries Joe... USA is not a good example.

Your argument is well known to me, and it is pernicious nonsense on several grounds. Being present at the testing may guard against outright tampering of the evidence, but it does little else. Some of the issues that I have raised depend up how the raw data are processed. The person doing the processing has made irrevocable decisions about the data; therefore, it represents his or her interpretation of the data set, not the actual data set itself. Once the raw data are processed, the processing parameters are set in stone, and I have previously given examples and quotes that explain why this is undesirable. However, the canard about the defense not being present utterly fails when one considers that some of the defense experts were brought in after the tests were completed. To the best of my knowledge, these include Dr. Tagliabracci, Dr. Hampikian, and Dr. Johnson. They had no opportunity to be present at the initial testing; therefore, there are even fewer grounds to deny them the raw data than the other experts. The appeal will document the failure of the prosecution to abide by sensible standards of discovery. I will discuss the appeal in a future post.

About Me

I am a biochemist who specializes in the chemical modification of proteins and the synthesis of potential enzyme inhibitors. I am particularly interested in the chemistry of phosphorus and sulfur as it can be applied to biochemical problems.